Identification of adiponutrin-related proteins as esterases and methods of use for the same

Abstract

The present invention provides methods of identifying polypeptides that have enzymatic activity associated with nutrient and/or energy homeostasis, and thus, are involved in the development of one or more cardiovascular and metabolic disorders, e.g., cardiovascular disease, obesity, insulin resistance, type 2 diabetes, dyslipidemia, nonalcoholic fatty liver disease, and metabolic syndrome. One such method comprises identifying a polypeptide as a member of the adiponutrin family of proteins. As such, the invention is related to the polynucleotides and polypeptides belonging to the adiponutrin family, and provides novel isolated and purified polynucleotides and polypeptides of a novel member of the adiponutrin family, patatin-like phospholipase domain 1 (PNPLA1). Also provided are methods of using the polynucleotides and polypeptides related to or provided by the invention for screening a test compound, e.g., a small molecule, antibody, etc., for the ability of the test compound to detect and/or modulate the activity of one or more members of the adiponutrin family of proteins. The present invention also is directed to novel methods for diagnosing, prognosing, and monitoring the progress of at least one cardiovascular and metabolic disorder using polynucleotides or polypeptides belonging to the adiponutrin family, and/or modulators of one or more members of the adiponutrin family. The present invention is further directed to novel therapeutics and therapeutic targets for the intervention (treatment) and prevention of cardiovascular and metabolic disorders arising from dysregulated energy homeostasis, as related to one or more members of the adiponutrin family of proteins.

1 . An isolated nucleic acid molecule having the nucleotide sequence of SEQ ID NO:22.  
     
     
         2 . An expression vector for inhibiting esterase activity, wherein the expression vector comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs:1, 4, 7, 10, 13, 16, 19, 22, 25, 52, 55, and portions thereof.  
     
     
         3 . A cell exhibiting an inhibited esterase activity, wherein the cell comprises the expression vector of  claim 2 .  
     
     
         4 . An expression vector for enhancing esterase activity, wherein the expression vector comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs:1, 4, 7, 10, 13, 16, 19, 22, 25, 52, 55, and portions thereof encoding active fragments of the polypeptides encoded by said SEQ ID NOs.  
     
     
         5 . A cell exhibiting an enhanced esterase activity, wherein the cell comprises the expression vector of  claim 4 .  
     
     
         6 . A method for identifying a modulator of the esterase activity of a member of the adiponutrin family of proteins, the method comprising the steps of contacting the member of the adiponutrin family with a candidate modulator and determining whether the candidate modulator modulates the esterase activity of the member of the adiponutrin family of proteins.  
     
     
         7 . The method of  claim 6 , wherein the esterase activity is lipid acyl hydrolase activity.  
     
     
         8 . The method of  claim 6 , wherein the esterase activity is lipase activity.  
     
     
         9 . A method for modulating esterase activity in a subject comprising the steps of: 
 selecting a subject in need of a modulator of the esterase activity of one or more members of the adiponutrin family of proteins; and    administering to the subject a therapeutically effective amount of such a modulator.    
     
     
         10 . A method for identifying a modulator of the enzymatic activity of a member of the adiponutrin family of proteins for the treatment of a disease selected from the group consisting of cardiovascular disease, obesity, insulin resistance, type 2 diabetes, dyslipidemia, nonalcoholic fatty liver disease, and metabolic syndrome, the method comprising determining whether a candidate modulator modulates the enzymatic activity of one or more members of the adiponutrin family of proteins.  
     
     
         11 . A method for treating a disease selected from the group consisting of cardiovascular disease, obesity, insulin resistance, type 2 diabetes, dyslipidemia, nonalcoholic fatty liver disease, and metabolic syndrome, the method comprising the steps of: 
 selecting a subject in need of a modulator of the enzymatic activity of one or more members of the adiponutrin family of proteins; and    administering to the subject a therapeutically effective amount of such a modulator.    
     
     
         12 . A pharmaceutical composition comprising a modulator of the activity of one or more members of the adiponutrin family of proteins and a pharmaceutically acceptable carrier.  
     
     
         13 . An expression vector for inhibiting lipase activity, wherein the expression vector comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs:1, 13, 52, and portions thereof encoding active fragments of the polypeptides encoded by said SEQ ID NOs.  
     
     
         14 . A cell exhibiting inhibited lipase activity, wherein the cell comprises the expression vector of  claim 13 .  
     
     
         15 . The method of  claim 6 , wherein the step of contacting the member of the adiponutrin family of protein with the candidate modulator comprises contacting a cell with the candidate modulator, wherein the cell comprises an expression vector comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:1, 4, 7, 10, 13, 16, 19, 22, 25, 52, 55, and portions thereof encoding active fragments of the polypeptides encoded by said SEQ ID NOs.  
     
     
         16 . The method of  claim 15 , wherein the cell is an adipocyte.  
     
     
         17 . The method of  claim 16 , wherein the adipocyte is a 3T3-L1 cell.  
     
     
         18 . The method of  claim 15 , wherein the step of contacting the cell with the candidate modulator comprises contacting the cell with isoproterenol.  
     
     
         19 . The method of  claim 15 , wherein the step of contacting the cell with the candidate modulator comprises contacting the cell with insulin.  
     
     
         20 . A method for identifying a modulator of a member of the adiponutrin family of proteins, the method comprising the step of contacting a cell with a candidate modulator, wherein the cell comprises an expression vector comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOs:1, 4, 7, 10, 13, 16, 19, 22, 25, 52, 55, and portions thereof encoding active fragments of the polypeptides encoded by said SEQ ID NOs.  
     
     
         21 . The method of  claim 20 , wherein the expression vector comprises the nucleic acid sequence of SEQ ID NO:1.  
     
     
         22 . The method of  claim 20 , wherein the cell is an adipocyte.  
     
     
         23 . The method of  claim 22 , wherein the adipocyte is a 3T3-L1 cell.  
     
     
         24 . The method of  claim 20 , wherein the step of contacting the cell with the candidate modulator comprises contacting the cell with isoproterenol.  
     
     
         25 . The method of  claim 20 , wherein the step of contacting the cell with the candidate modulator comprises contacting the cell with insulin.